skip to main content

DOE PAGESDOE PAGES

Title: Structural and functional analysis of the GABARAP interaction motif (GIM)

Through the canonical LC3 interaction motif (LIR), [W/F/Y]–X 1–X 2[I/L/V], protein complexes are recruited to autophagosomes to perform their functions as either autophagy adaptors or receptors. How these adaptors/receptors selectively interact with either LC3 or GABARAP families remains unclear. Herein, we determine the range of selectivity of 30 known core LIR motifs towards individual LC3s and GABARAPs. From these, we define a GABARAP Interaction Motif (GIM) sequence ([W/F]–[V/I]–X 2–V) that the adaptor protein PLEKHM1 tightly conforms to. Using biophysical and structural approaches, we show that the PLEKHM1–LIR is indeed 11–fold more specific for GABARAP than LC3B. Selective mutation of the X 1 and X 2 positions either completely abolished the interaction with all LC3 and GABARAPs or increased PLEKHM1–GIM selectivity 20–fold towards LC3B. Finally, we show that conversion of p62/SQSTM1, FUNDC1 and FIP200 LIRs into our newly defined GIM, by introducing two valine residues, enhances their interaction with endogenous GABARAP over LC3B. In conclusion, the identification of a GABARAP–specific interaction motif will aid the identification and characterization of the expanding array of autophagy receptor and adaptor proteins and their in vivo functions.
Authors:
 [1] ;  [2] ;  [3] ;  [4] ;  [5] ;  [1] ;  [1] ;  [6] ;  [1] ; ORCiD logo [7] ; ORCiD logo [8] ; ORCiD logo [9]
  1. Goethe Univ., Frankfurt am Main (Germany)
  2. Goethe Univ. School of Medicine, Frankfurt (Main) (Germany)
  3. Univ. of Canterbury, Christchurch (New Zealand)
  4. Univ. of Dundee, Dundee (United Kingdom)
  5. Univ. of Canterbury, Christchurch (New Zealand); High Energy Accelerator Research Organization (KEK), Tsukuba Ibaraki (Japan)
  6. High Energy Accelerator Research Organization (KEK), Tsukuba Ibaraki (Japan); SLAC National Accelerator Lab., Menlo Park, CA (United States); Structural Biology (School of Medicine), Stanford, CA (United States)
  7. Goethe Univ. School of Medicine, Frankfurt (Main) (Germany); Goethe Univ., Frankfurt am Main (Germany)
  8. Univ. of Canterbury, Christchurch (New Zealand); Univ. of Melbourne, Parkville, VIC (Australia)
  9. Goethe Univ. School of Medicine, Frankfurt (Main) (Germany); Univ. of Dundee, Dundee (United Kingdom)
Publication Date:
Grant/Contract Number:
AC02-76SF00515; SFB 1177; EXC115; SPP 1580; HFSP RGP55; UOCX1208; W911NF‐11‐1‐0481; UOC1013; UOC1506; FY2012; Az. 10.14.2.208; 202061/Z/16/Z; T16/44
Type:
Accepted Manuscript
Journal Name:
EMBO Reports
Additional Journal Information:
Journal Volume: 18; Journal Issue: 8; Journal ID: ISSN 1469-221X
Publisher:
European Molecular Biology Organization
Research Org:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org:
USDOE
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; autophagy; Atg8; GABARAP; LC3; PLEKHM1
OSTI Identifier:
1390293